1 / 27

A systems approach to study plant disease resistance mechanisms

A systems approach to study plant disease resistance mechanisms. Fumiaki Katagiri Dept. of Plant Biology Univ. of Minnesota Microbial and Plant Genomics Institute. Oct. 17, 2005 CBCB seminar Cargill Bldg. Acknowledgements. Stanford Functional Genomics Facility. Univ. of Minnesota

belva
Télécharger la présentation

A systems approach to study plant disease resistance mechanisms

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. A systems approach to study plant disease resistance mechanisms Fumiaki Katagiri Dept. of Plant Biology Univ. of Minnesota Microbial and Plant Genomics Institute Oct. 17, 2005 CBCB seminar Cargill Bldg.

  2. Acknowledgements Stanford Functional Genomics Facility Univ. of Minnesota Masanao Sato Duke University Dong Wang Natalie Weaver Xinnian Dong Raka Mitra Jane Glazebrook Supercomputing Institute Zheng Jin Tu Carnegie Institution Shauna Somerville High Throughput Biological Analysis Facility Nick Hahn Harvard Medical Sch. Julia Dewdney Fred Ausubel Funding:

  3. Inducible Defense of Plants Effectors Pathogen Recognition Signal Transduction

  4. Arabidopsis plants infected with Pseudomonas syringae carrying avrRpt2 rps2- RPS2+

  5. How should we study a signaling network? INPUTS Black Box OUTPUTS If we only observe inputs and outputs, we can never specify the mechanism in the black box, however many observations we make.

  6. How should we study a signaling network? INPUTS Black Box Readouts from the inside So, we beat it up (perturbations) and try to see what changes occur inside the box (i.e., network).

  7. Our strategy • T-DNA insertion lines for genetic perturbations. • A small scale microarray for a wide spectrum, quantitative measurements.

  8. “Mini”-array • Long oligo (50-70 mers) spotted array • 464 pathogen-responsive genes – representing diverse expression patterns • 107 normalization genes for array-to-array normalization – representing a wide range of expression levels • 5 spiking oligos for quantitation check • Tracking oligo for pixel-to-pixel calibration • Locally factorial, globally overlapping design, which allows use of a global linear model.

  9. Locally factorial, globally overlapping design In each tile, 72 probes x 2 are common in each of the subarrays. A single subarray has 144 probes x 2 = 288 spots

  10. Technical reproducibility (Mean, 1 pen x duplicates) R= 0.945 slope = 0.94 y-intercept = -0.19 R = 0.984 slope = 1.00 y-intercept = -0.02

  11. Technical reproducibility of Miniarray (Mean, 4 pens x duplicates) R= 0.973 slope = 0.96 y-intercept = -0.32 R = 0.990 slope = 1.06 y-intercept = 0.40

  12. Statistical model for the miniarray expression value (1)- linear model Indices: i, probe (gene); j, pen (subarray); r, replicate Sijr, Log-transformed measured value (median of ratios) Ai, probe contribution; Bj, pen contribution; Cr, replicate contribution; Ejr, pen-replicate interaction; eijr, error

  13. Technical reproducibility of Miniarray (linear model) R= 0.978 slope = 0.95 y-intercept = -0.12 R = 0.996 slope = 1.04 y-intercept = 0.20

  14. Statistical model for the miniarray expression value xj yj jth subarray Indices: i, probe (gene); j, pen (subarray); r, replicate Sijr, Log-transformed measured value (median of ratios) Ai, probe contribution; Bj, pen contribution; Cr, replicate contribution; Ejr, pen-replicate interaction; fj(xj, yj), spatial correction within jth subarray; eijr, error

  15. Technical reproducibility of Miniarray (linear model + spatial smoothing function) R= 0.982 slope = 0.96 y-intercept = -0.14 R = 0.994 slope = 1.06 y-intercept = 0.37

  16. Distribution of spot-by-spot errors The mean of measured values as the expression value for each gene (variance = 0.18) After correction with a linear model only (variance = 0.13) After correction with a linear model and a spatial smoothing function (variance = 0.062) buf24hx1

  17. Spatial error distribution(same color scale) Mean Linear model Linear model + Spatial smoothing function psm24hx1

  18. Correlation in the expression ratio between miniarray and Affy data (linear model + spatial smoothing function) R = 0.846 slope = 1.08 y-intercept = 0.37

  19. The major cause of the discrepancy is a limited dynamic range of the miniarray at the low end At least one of the miniarray exp values is < -3.5 Both values are > -3.5 Exp value range (log2-transformed) : -5.2 to 6.6

  20. Collaboration stylein systems biology research

  21. What’s systems biology?- My definition • Study the topology and dynamics of biological networks that underlie biological phenomena. • It is not just systematic collection of biological data.

  22. International study ofsystems biology research • Panel members: Marvin Cassman (Chair), Adam Arkin (UC Berkeley), Frank Doyle (UCSB), Doug Lauffenburger (MIT), Cindy Stokes (Enteros), Fumi Katagiri (U of Minn) • Organized by World Technology Evaluation Center (http://wtec.org). • Sponsored by NSF, DOE, DARPA, NASA, NCI, NIBIB, NIST, and EPA.

  23. Computational/ Theoretical Experimental Relatively little instructed global profiling and/or systematic approaches Network inference Early stage Other pre-existing data Validation, Refinement, Parameters, … Highly instructed, focused experiments Modeling Later stage Prediction, Demand, …

  24. Old style - Division of labor Theoreticians/ Computer scientists Experimentalists Areas of expertise

  25. Desired style – Highly interactive Theoreticians/ Computer scientists Experimentalists Areas of expertise

  26. An alternative Theoreticians/ Computer scientists Experimentalists Areas of expertise

  27. http://www.cbs.umn.edu/class/fall2005/pbio/5960-2/

More Related